Tissue factor procoagulant activity in the tumor cell lines and plasma of dogs with various malignant tumors

Figures

• 

  Fig. 1.. Setting of flow cytometry to detect microparticles (MPs). MPs were defined as both events of 0.5–0.9 µm diameter and Annexin V-positive events. (A) Bead subsets (0.5, 0.9, and 3.0 µm beads in diameter) were first recognized based on their side scatter (SS) and FL1 fluorescence properties. (B) In the forward scatter (FS) histogram with distributions of 0.5 and 0.9 µm beads, the threshold level of FSC was set to achieve a percentage of 49.0–51.0% for 0.5-µm beads (M-0.5µm). (C) On an FSC log ×SSC log cytogram, the MP gate was considered a region under 0.9 µm beads. (D) A representative result in Ud2 is shown. The region below the solid red line and the shaded region in the histograms indicated staining with and without (negative control) PerCP-eFluor® 710 dye-conjugated Annexin V, respectively.

• 

  Fig. 2.. Expression of tissue factor (TF) on canine tumor cell lines. Cell surface expressions of TF were analyzed via flow cytometry; (A) D17 (negative control) and T24 (positive control), (B) mammary gland tumor, (C) hemangiosarcoma, (D) malignant melanoma, and (E) lymphoma cell lines. The shaded region and the region below the solid line in the histograms indicated staining with isotype control and anti-TF antibody, respectively. Results shown are representative of a minimum of three independent experiments.

• 

  Fig. 3.. Numbers of microparticles (MPs) in the culture supernatants of canine tumor cell lines. MPs in the culture supernatants of canine mammary gland tumor, hemangiosarcoma, malignant melanoma, and lymphoma cell lines were measured via flow cytometry. The MP counts are shown as mean values and standard deviation. Results shown are representative of at least three independent experiments.

• 

  Fig. 4.. Tissue factor procoagulant activity (TF-PCA) of cellular surface and microparticles (MPs) in the culture supernatants of canine tumor cell lines. TF-PCA of the cell surface and MPs in the culture supernatants of canine mammary gland tumor, hemangiosarcoma, malignant melanoma, and lymphoma cell lines are shown. D17 and T24 cells were used as negative and positive controls, respectively. TF-PCA was presented as mean values and standard deviation. Results shown are representative of at least three independent experiments.